Objective To investigate the expressions of AKT and ERK1/2 proteins in hypopharyngeal squamous cell carcinoma and normal hypopharyngeal mucosa. Methods The expressions of AKT and ERK1/2 proteins were exam-ined by immunohistochemical S-P technique in 60 patients with hypopharyngeal squamous cell carcinoma and 15 cases of normal hypopharyngeal mucosa . The relationship between expressions of AKT and ERK1/2 proteins and clinical pathologi-cal feathers was analyzed. Results The positive rates of the expressions of AKT and ERK1/2 were 78.3% (47/60) and 66.7%(40/60) in 60 cases of hypopharyngeal squamous cell carcinoma, which were significantly higher than those in 15 cas-es of normal hypopharyngeal mucosa [13.3%(2/15) and 6.7%(1/15), P<0.05]. The lower the degree of differentiation, the later the clinical stage, the higher the positive expression rates of AKT and ERK1/2. There were significantly higher expres-sions of AKT and ERK1/2 in patients with lymph node metastasis than those of patients without lymph node metastasis (P<0.05 or P<0.01). There was a positive correlation between expression levels of AKT and ERK1/2 (rs=0.400,P<0.05). Con-clusion There were higher expression levels of AKT and ERK1/2 in hypopharyngeal squamous cell carcinoma. The activa-tion of AKT and ERK1/2 proteins promotes hypopharyngeal occurrence, development and metastasis.

Objective:To detect the biological activity of the expression products of the human ?-NGF cDNA in CHO cell line and purified the seperated protein.Methods:Human ?-NGF was transfected with lipofectamine reagent into CHO cell line.The biological activity was analyzed by objection with microscope and the method of MTT.The pure protein was proved by SDS-PAGE analysis.Results:The protein in the culture supernatant of the positive CHO cells transfected with ?-NGF gene could stimulate the growth of PC12 cell line and go into BBB.Conclusion:The target gene expressed successfully in the transfected CHO cell line and had good biological activity. [

0.05).Internal fixation could make the adjacent segment rigidity degrade and stress raise significanly(P

Aim To construct the eukaryotic expression vector of hIL-24 cDNA,and express it in CHO cells and detect its anti-tumor effect of recombinant hIL-24 protein.Methods Constructed pcDNA3-hIL-24 was identified by endonucleases digestion & PCR.The recombinant expression plasmids were transfected into CHO cells,human hIL-24 expressed in CHO cells was detected with RT-PCR.The apoptosis-inducing activities of recombinant protein hIL-24 was tested by MTT assay,Hoechst& FCM assay,and the expression of IL-6 and IFN-? from PBMC induced by rhIL-24 was tested by ELISA.Results The eukaryotic expression vector pcDNA3-hIL-24 was constructed correctly.Stable expression of human IL-24 in CHO cells was identified with RT-PCR.The apoptosis of A549 cells induced by hIL-24 was proved by Hoechst & FCM assay,and the expression of IL-6 and IFN-? from PBMC induced by rhIL-24 was identified with ELISA.Conclusion The successful stable expression & experimental study of apoptosis effect of human IL-24 gene lay the foundation for the further study of molecular mechanism of hIL-24 on anti-tumors and potential application.

Objective:To construct the cells which can express LIF protein steadily and study the effect of rhLIF and IL-24 on HL-60 cells.Methods:ECV-304 cells were infected by eukaryotic expression plasmid pcDNA3-LIF,then the positive cells were selected by G418.The positive cells were collected and their culture supernatant was stored for further use.Expression of LIF mRNA was detected by RT-PCR.The pAdeasy-1-pAdTrack-CMV-IL-24 was extracted from DH5a.The recombined adenovirus vector was lineared with PacI and transfected into 293 cells.The IL-24 recombined adenovirus(Ad-IL-24) was obtained and used to infect HL-60 cells.At the same time,the culture supernatant containing rhLIF was added into the HL-60 cells which was identified as positive expression of IL-24 by RT-PCR.Synergistic effect of the cytokines on HL-60 cells was tested by LSCM, FCM and immunohistochemistry stain assay.Results:The cells expressing LIF protein steadily were constructed successfully, the high titer of the recombined adenovirus(Ad-IL-24) was obtained. Expression of IL-24 in infected HL-60 cells was identified by RT-PCR.The apoptosis of HL-60 cells induced by rhLIF and IL-24 was proved by LSCM ,FCM and immunohistochemistry stain assay. They had synergistic effect.Conclusion:rhLIF and IL-24 can inhibit growth of HL-60 cells and induce apoptosis of the cells.They have synergistic effect.

Objective:To construct a recombinant prokaryotic expression vector pGEX-5X-3/hIL-17F and express it in E.coli and to explore the biological activities of human IL-17F fusion protein.Methods:The coding sequence of the mature human IL-17F(minus the signal peptide) was amplified from pUCm-T/hIL-17F by PCR and subcloned into the prokaryotic expression vector pGEX-5X-3 to express glutathione S-transferase(GST) fusion protein. The fusion protein was induced in E.coli BL21 by IPTG and purified by standard methods reported in prokaryotic system. The purified GST-hIL-17F fusion protein was identified by Western blot. The proliferation of ECV304 cells was observed by incubating them with soluble GST-hIL-17F fusion protein by MTT assay. The concentrations of IL-6, IFN-? and TNF-? in the supernatants of ECV304 cells were determined by ELISA. The effect of GST-hIL-17F on the angiogenesis of the chick chorioallantoic membrane was assessed by CAM assay.Results:A 41 kD fusion protein was effciently induced in E.coli BL21 by IPTG, accounting for about 55% of the total bacterial protein. The purified GST-hIL-17F fusion protein was identified by Western blot. GST-hIL-17F fusion protein had obvious biological activity to inhibit the proliferation of ECV304 cells and enhance IL-6 secretion. GST-hIL-17F had a marked antiangiogenic activity.Conclusion:The preliminary study of hIL-17F recombinant prokaryotic expression and its antiangiogenic effect has been successful, which lays a foundation for future research on the mechanism of antiangiogenesis and clinical application of recombinant hIL-17F protein.

Objective To explore the clinical effents of bevacizumab on the recrudescence of the limbal corneal epithelial cell auto-graft transplantation in treating patients with pterygium.Methods The clinical data of 75 cases(99 eyes) with pterygium were retraspectively reviewed,and they were divided into 3 groups by different conservative treatment.A group:the limbal corneal epithelial cell auto-graft transplantation combined with bevacizumab.B group:the limbal corneal epithelial cell auto-graft transplantation combined with MMC.C group:the limbal corneal epithelial cell auto-graft transplantation.After follow-up for 3 months,the curative effect and recurrence were compared between the two groups.Results The recurrence of three groups was significantly different( x2 =12.267,P ＜ 0.05 ).The reccurrence rate of A,B,C group were 12.1％,15.2％,45.5％.The recurrence rate of A group and B group wasn't significantly different(x2 =2.117,P ＞0.05).The recurrence rate of B group and C group wasn't statistically different( x2 --3.930,P ＜ 0.05 ).The recurrence rate of A group and C group was significantly different( x2 =4.155,P ＜ 0.05 ).After 1 week,all patients had different degrees of eye pain,photophobia or tearing,and disappeared after 1 week;2 patients in group B found that limbal shallow scleral necrosis,superficial punctate keratitis.The average time of removal of stitches in group A was 5.9d,group B was 7.0d and group C was 7.5d.Conclusion Bevacizumab could obviously reduce the recrudescence of the limbal corneal epithelial cell auto-graft transplantation in treating patients with pterygium.It was safe with less complications and good prognosis.It was worthy for being widely used in treatment of pterygium.

Objective To construct the human IL-24 recombined adenovirus(Ad-IL24)and to observe the effect of Ad-IL24 on ex vivo culture of HL-60 cells.Methods pAdTrack-CMV-IL24 was constructed by PCR with pcDNA3.0-IL24 recombined plasmid as a template,enzyme digestion and ligation.The pAdTrack-CMV-IL24 lineared by Pme Ⅰ was co-transformed into BJ5183 with pAdEasy-1.The pAdEasy-1-pTrack-CMV-IL24 recombined adenovirus vector was lineared with Pac Ⅰ and then transfected in to QBI-293A cells.The Ad-IL24 was obtained and used to infect HL-60 ceils,and the effect on HL-60 cells was tested by LSCM,Mrrr,FCM,ELISA and immunohistochemistry staining.Results The pAdEasy-1-pTrack-CMV-IL24 vector was constructed and the Ad-IL24 was successfully obtained.The effect of inhibiting growth of HL-60 cells and inducing cell apoptosis by IL-24 was proved by LSCM,MTT and FCM.IL-24 down-regulated the expression of anti-apoptosis factor Bcl-2 and increased the secretion of IFN-γ,TNF-α of HL-60 cells.Conclusion Ad-IL24 can inhibit the growth of HL-60 cells and induce cell apoptosis through down-regulating expression of anti-apoptosis factor and increasing the secretion of IFN-γ,TNF-α of HL-60 cells.The human Ad-IL24 may provide a basic study for the future target therapy to tumors.

To study the current situation and affecting factors of bilingual teaching for Neurology,we investigated the students of a five-year medical undergraduade in clinical medicine with a questionaire.It will provide the data and reference to effectively improve bilingual teaching program for Neurology.

Objective To promote the standardization of medical practice in China by comparing the discrepancies of diagnosis and medication strategies for benign prostatic hyperplasia (BPH) between urologists and geriatricians.Methods Departments of urology and geriatrics in general hospitals in China were selected through stratified sampling and 145 315 patients who went for the outpatient service in certain days during December 2011 to December 2012 were recruited by cluster.Questionnaires were completed by corresponding doctors and data of patient clinical characteristics as well as diagnostic and therapeutic pattern were collected.Results A total of 142 511 valid questionnaires were collected with 119 426 from urology and 23 085 from geriatrics.The average age of BPH patients was (68.34±8.61) years and the average IPSS was 17.93±4.73.The rates of diagnostic method were IPSS (99.2％),ultrasonic inspection (86.0％),digital rectal examination (DRE) (66.0％) and urine flow rate test (55.2％).The rate of ultrasonic application was lower while the rates of other methods were higher among urologists (P＜0.01) ; the rate of DRE among urologists was 2.2 times to that among geriatricians.For patients with moderate or severe symptoms,most urologists (72.1％ in moderate patients and 79.9％ in severe patients) applied drug-combination strategy and the rate was 2 times to that among geriatricians (P＜0.01).In contrast,most geriatricians (59.7％ in moderate patients and 56.1％ in severe patients) applied single drug to BPH patients.Urologists utilized α-receptor blockers,plant preparations and traditional Chinese medicine more frequently than geriatricians (P＜0.01).Conclusions Urologists performed more integrated diagnosis strategy compared with geriatricians.The urologists tended to use 5α-reductase inhibitor combined with α-receptor blocker to treat BPH patients with moderate or severe symptoms,while geriatricians preferred 5α-reductase inhibitor alone.